Intracellular localization of tyrosine kinase substrates beneath crossfnked surface immunoglobulins in B cells

Shuji Takagi, Masanori Daibata, Thomas J. Last, Robert E. Humphreys, David C. Parker, Takeshi Sairenji

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Crosslinking of surface immunoglobulins (sIg) in B cells led to the accumulation of submembranal phosphotyrosine, which was followed morphologically with the PY20 antiphosphotyrosine monoclonal antibody. Phosphotyrosine was not detected before sIg crosslinking. After sIg crosslinking, phosphotyrosine-containing proteins were redistributed from scattered small clusters near the plasma membrane to ajuxtanuclear region, where immunofluorescent staining decreased with time. Double immunofluorescent staining of individual cells showed accumulation of phosphotyrosine beneath crosslinked sIg molecules at the cell surface. The sIg molecules were subsequently internalized more rapidly than the phosphotyrosine-containing molecules were redistributed. Genistein, a protein tyrosine kinase (PTK) inhibitor, blocked intracellular tyrosine phosphorylations but not cell surface patching of crosslinked sIg. When polyacrylamide beads coated with anti-Ig antibodies were added to the cells, intracellular tyrosine phosphorylation occurred beneath the regions of contact with the beads. This study provides an independent line of evidence confirming recent biochemical experiments that show that crosslinking of the antigen receptor induces PTK activity in B cells, and that components of the newly described sIg’complex are among the PTK substrates. The surprising finding that the bulk of the induced phosphotyrosine remains associated with crosslinked s1g for many minutes suggests a role for complex local protein interactions in phosphotyrosine-mediated signal transduction through the antigen receptor of B cells.

Original languageEnglish (US)
Pages (from-to)381-388
Number of pages8
JournalJournal of Experimental Medicine
Volume174
Issue number2
DOIs
StatePublished - Aug 1 1991

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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